CN113831548A - Application of penicillium MJ51 extracellular fluid in preparation of water-soluble humic acid by dissolving lignite - Google Patents
Application of penicillium MJ51 extracellular fluid in preparation of water-soluble humic acid by dissolving lignite Download PDFInfo
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Abstract
The invention provides a novel method for preparing water-soluble humic acid by dissolving lignite with penicillium MJ51 extracellular fluid. Firstly, lignite is crushed and sieved, the lignite is dissolved by adopting a cell-free culture solution of penicillium MJ51, and filtrate is collected by filtration to obtain an extracting solution containing humic acid. Compared with humic acid extracted by a traditional strong alkali method, the water-soluble humic acid solution prepared by the method has neutral pH, higher nitrogen content, stronger flocculation resistance and higher content of oxygen-containing functional groups, and can be directly used as a water-soluble humic acid fertilizer. The method has the advantages of simple process, high efficiency, environment-friendly biological solvent for extracting humic acid, high physiological activity of humic acid products and capability of solving the problems of complicated flow and environmental pollution in the traditional process, and the extraction of the humic acid in the lignite by the penicillium MJ51 extracellular fluid provides a new way for the efficient utilization of the lignite and the application of the humic acid products in agriculture, and the preparation method has good development potential.
Description
Technical Field
The invention relates to the technical field of coal processing, in particular to application of penicillium MJ51 extracellular fluid in preparation of water-soluble humic acid by dissolving lignite.
Background
The lignite is low-rank coal with low coalification degree and high reaction activity. Lignite has high moisture content and low calorific value, and the direct combustion efficiency is low and pollutes the environment, so that it is necessary to develop a new technology for utilizing lignite, which is economically efficient and environmentally safe. At present, the extraction of humic acid from lignite is an important way for realizing the conversion and utilization of non-fuel and high added value of lignite.
At present, alkali extraction method, acid extraction method, microorganism dissolution method, organic solvent extraction method and the like are reported as methods for extracting humic acid from lignite, wherein the alkali extraction method is a classical method for extracting humic acid. The acid oxygen-containing functional groups of the humic acid, such as carboxyl, phenolic hydroxyl and the like, firstly react with alkali substances to generate soluble humate, then acid is added to adjust the pH value to be less than 2, and solid-liquid separation is carried out, so that the purposes of purifying and separating the humic acid are achieved. However, the solution is strong in alkalinity when being directly used, a large amount of energy is consumed when the solution is dried, the solution is precipitated by acid, washed and precipitated by water and then dried, high-purity humic acid can be obtained, the process flow is complex, a large amount of acid is consumed, the production efficiency is low, and the strong base solvent and the organic solvent used in the process of extracting the humic acid are not green environment-friendly solvents, so that the environmental pollution is easily caused. In addition, the humic acid product prepared by the alkali extraction method has the defects of few active groups, being not beneficial to the subsequent utilization of saline-alkali soil and the like. Aiming at the conditions, the invention provides a humic acid extraction process which is simple in process and environment-friendly, namely, the microbial extracellular fluid is used for replacing a strong alkali reagent to extract humic acid in lignite.
It has been reported that microorganisms can dissolve lignite by secreting alkaline substances, thereby promoting the release of water-soluble humic acid in lignite. Compared with the conventional alkali extraction method, after the lignite is dissolved by the microorganisms, the water-soluble humic acid content is improved, and the humic acid content is increasedThe structures of (A) are also different and mainly show that the molecular weight and the aromatic condensation degree are reduced, the nitrogen content is increased, and the biological activity is enhanced. Various microorganisms such as Penicillium sp.P6, Bacillus sp.Y7, Streptomyces fulvisimus K59, etc. have been reported to have the ability to produce alkaline secondary metabolites and this alkaline substance plays an important role in solubilizing lignite to produce highly physiologically active humic acid. The thermophilic alkaline substance secreted by Bacillus sp.Y7 can dissolve 36.77% of lignite within 12d (Jiang, F., Li, Z., Lv, Z., Gao, T., Yang, J., Qin, Z., Yuan, H.,2013.The biochemical of lignite by Bacillus sp.Y7 and chromatography of The soluble products. Fuel,103, 639-645.). Streptomyces fulvisimus K59 also produces alkaline substances to dissolve lignite, which is subjected to 5N HNO3After pretreatment, the biological solubilization effect is optimal, and the concentration of humic acid in the solubilized product is 15 times of that of raw coal (Sobolczyk-Bednarek, J., Choi ń ska-Pulit, A.,
W.,2021.Biosolubilization of low-rank coal by the newly isolated strain Streptomyces fulvissimus K59.Fuel,301,121082.)。
compared with bacteria, the fungi can play a stronger role in dissolving the lignite to change the structure of the humic acid product and improve the physiological activity of the humic acid. The penicillium species in the fungus have the ability to produce alkaloids, which have the function of dissolving lignite. Some strains of Penicillium have been shown to solubilize lignite by secretion of alkaline substances, surfactants, extracellular enzymes (Achi, 1994; Yuan et al, 2006). The prior art discloses a strain of penicillium MJ51, and a culture of the penicillium MJ51 can be used for producing fulvic acid by degrading lignite. However, no report is provided about the penicillium MJ51 extracellular fluid in the preparation of water-soluble humic acid by dissolving lignite.
Disclosure of Invention
The invention aims to provide a new way for preparing water-soluble humic acid by dissolving lignite with penicillium MJ51 extracellular fluid.
In order to achieve the above object, the present invention provides the following technical solutions:
the invention provides application of penicillium MJ51 extracellular fluid in preparation of water-soluble humic acid by dissolving lignite.
Preferably, the method for preparing water-soluble humic acid by dissolving lignite comprises the following steps:
mixing the penicillium MJ51 extracellular fluid with oxidized lignite to obtain a mixed solution, and dissolving coal in the mixed solution at the temperature of 60-90 ℃ to obtain a dissolved solution, wherein the dissolved solution contains water-soluble humic acid.
Preferably, the mass concentration of the oxidized lignite in the mixed solution is 3-5%.
Preferably, the preparation method of the oxidized lignite comprises the following steps: mixing nitric acid and lignite, and then carrying out oxidation-reduction reaction;
and carrying out solid-liquid separation on the reaction feed liquid obtained by the oxidation-reduction reaction to obtain a solid component, wherein the solid component is oxidized lignite.
Preferably, the temperature of the oxidation-reduction reaction is 25-30 ℃.
Preferably, the equivalent concentration of the nitric acid is 4-6N.
Preferably, the preparation method of the penicillium MJ51 extracellular fluid comprises the following steps:
inoculating the spore suspension of the penicillium MJ51 into a liquid culture medium, and culturing to obtain a culture solution;
isolating the extracellular fluid of penicillium MJ51 from the culture medium;
the spore concentration of the penicillium MJ51 in the spore suspension is (1-9) multiplied by 107A plurality of; the volume ratio of the spore suspension to the culture medium is 5: (80-120).
Preferably, the temperature of the culture is 25-30 ℃; the culture time is 55-65 h.
Preferably, the liquid culture medium takes water as a solvent and comprises the following components in concentration: 12-18 g/L, KNO of sucrose32～4g/L、KH2PO4 0.8～1.2g/L、Na2HPO4 0.3～0.8g/L、MgSO40.4-0.6 g/L; the pH value of the culture medium is 5.8-6.2.
Preferably, the penicillium MJ51 has the deposit number: CGMCC No. 18120.
The invention provides a novel method for preparing water-soluble humic acid by dissolving lignite with penicillium MJ51 extracellular fluid. Firstly, lignite is crushed and sieved, the lignite is dissolved by adopting a cell-free culture solution of penicillium MJ51, and filtrate is collected by filtration to obtain an extracting solution containing humic acid. Compared with humic acid extracted by a traditional strong alkali method, the water-soluble humic acid solution prepared by the method has neutral pH, higher nitrogen content, stronger flocculation resistance and higher content of oxygen-containing functional groups, and can be directly used as a water-soluble humic acid fertilizer. The method has the advantages of simple process, high efficiency, environment-friendly biological solvent for extracting humic acid, high physiological activity of humic acid products and capability of solving the problems of complicated flow and environmental pollution in the traditional process, and the extraction of the humic acid in the lignite by the penicillium MJ51 extracellular fluid provides a new way for the efficient utilization of the lignite and the application of the humic acid products in agriculture, and the preparation method has good development potential.
Drawings
FIG. 1 is a structural characterization of humic acid, where (a) is FTIR spectrum of HA sample and (b) is solid state cross-polarization magic angle rotation13C nuclear magnetic resonance spectrum, wherein HA-control is humic acid extracted from coal by sodium hydroxide; HA-CFFs are humic acids extracted from coal by cell-free filtrate (CFFs).
Biological preservation Instructions
The blue mold MJ51, Latin article is Penicillium camemberti, is preserved in China general microbiological culture Collection center in 20.08.2019, the preservation address is No. 3 of West Lu No.1 of the sunward area of Beijing, and the preservation number is CGMCC No. 18120.
Detailed Description
The invention provides application of penicillium MJ51 extracellular fluid in preparation of water-soluble humic acid by dissolving lignite.
In the invention, the structure of the penicillium MJ51 extracellular fluid can be changed, the aliphatic carbon and carboxyl carbon contents in the water-soluble humic acid structure can be increased, the aromatic carbon content can be reduced, and the water-soluble humic acid extracted from the penicillium MJ51 extracellular fluid has lower molecular weight, higher nitrogen content and flocculation limit value.
In the present invention, the Penicillium MJ51(Penicillium camemberti) has the deposit number: CGMCC No. 18120.
In the present invention, the method for preparing the penicillium MJ51 extracellular fluid preferably comprises the following steps: inoculating the spore suspension of the penicillium MJ51 into a liquid culture medium, and culturing to obtain a culture solution; isolating the extracellular fluid of penicillium MJ51 from the culture medium; the spore concentration of the penicillium MJ51 in the spore suspension is preferably (1-9) multiplied by 107A plurality of; the volume ratio of the spore suspension to the culture medium is preferably 5: (80-120), more preferably 1: 100. in the invention, the temperature of the culture is preferably 25-30 ℃; the culture time is preferably 55-65 h, and more preferably 60 h. In the present invention, the culture mode is preferably shake culture; the rotation speed of the shaking culture is preferably 150-200 rpm, and more preferably 160-180 rpm.
In the invention, the liquid culture medium takes water as a solvent and comprises the following components in concentration: 12-18 g/L, KNO of sucrose3 2～4g/L、KH2PO4 0.8～1.2g/L、Na2HPO4 0.3～0.8g/L、MgSO40.4-0.6 g/L; the pH value of the culture medium is 5.8-6.2; the pH value of the culture medium is preferably 5.8-6.2, and more preferably 6. In the invention, the liquid culture medium takes water as a solvent and comprises the following components in concentration: sucrose 15g/L, KNO3 3g/L、KH2PO4 1g/L、Na2HPO40.5g/L and MgSO4 0.4～0.6g/L。
In the present invention, the lignite is preferably high-rank lignite; the particle size of the lignite is preferably 0.18-0.25 mm, and more preferably 0.2 mm.
In the invention, the method for preparing humic acid by dissolving lignite comprises the following steps: mixing the penicillium MJ51 extracellular fluid with oxidized lignite to obtain a mixed solution, and dissolving coal in the mixed solution at the temperature of 60-90 ℃ to obtain a dissolved solution, wherein the dissolved solution contains water-soluble humic acid.
In the present invention, the mass concentration of the oxidized lignite in the mixed liquor is preferably 3% to 5%, more preferably 4%.
In the present invention, the method for producing oxidized lignite preferably comprises the steps of: the preparation method of the oxidized lignite comprises the following steps: mixing nitric acid and lignite, and then carrying out oxidation-reduction reaction; and carrying out solid-liquid separation on the reaction feed liquid obtained by the oxidation-reduction reaction to obtain a solid component, wherein the solid component is oxidized lignite. In the invention, the temperature of the oxidation-reduction reaction is preferably 25-30 ℃. In the present invention, the equivalent concentration of the nitric acid is preferably 2 to 6N, and more preferably 4N.
In the invention, the ratio of the volume of the nitric acid to the mass of the lignite is preferably (20-30) mL: 1g, more preferably 25 mL: 1g of a compound; the temperature of the oxidation-reduction reaction is preferably 25-30 ℃, and more preferably 28 ℃; the time of the oxidation-reduction reaction is preferably 40-50 h, and more preferably 48 h. In the present invention, the solid-liquid separation is preferably performed by centrifugation; the rotating speed of the centrifugation is preferably 6000-8000 rpm, and more preferably 7000 rpm; the time for centrifugation is preferably 8-12 min, and more preferably 10 min. The present invention preferably further comprises, after collecting the solid component, washing the solid component until the wash liquor is colorless and has a pH > 5; the reagent used for washing is preferably deionized water; after the washing, preferably, the method further comprises drying the washed components; the drying mode is preferably drying; the drying temperature is preferably 70-90 ℃, and more preferably 80 ℃; the drying time is preferably 30-40 h, and more preferably 36 h.
After the dissolved solution is obtained, the method preferably further comprises the step of separating humic acid from the dissolved solution, and the method comprises the following steps: and filtering the dissolved solution, collecting filtrate, adjusting the pH value of the filtrate to 1.8, standing, centrifuging, and collecting precipitate to obtain humic acid. In the present invention, the filter paper used for the filtration is preferably Whatman No. 1; the reagent used for adjusting the pH value of the filtrate is preferably 6.0MHCl aqueous solution; the standing time is preferably 20-30 h, and more preferably 24 h; the rotation speed of the centrifugation is preferably 8000-12000 rpm, and more preferably 10000 rpm; the time for centrifugation is preferably 3-8 min, and more preferably 5 min. After the precipitate is collected, the invention preferably also comprises washing the precipitate and drying; the reagent used for cleaning is preferably distilled water; the temperature of the drying is preferably 60 ℃; the drying time is based on drying to constant weight. In the present invention, the preservation temperature of the humic acid is preferably 4 ℃.
The technical solution of the present invention will be clearly and completely described below with reference to the embodiments of the present invention. It is to be understood that the described embodiments are merely exemplary of the invention, and not restrictive of the full scope of the invention. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
The lignite samples adopted in the embodiment and the comparative example are collected from Yunnan Miller, are crushed, ground and sieved to prepare raw coal samples with the particle size of 0.18-0.25 mm, and are dried at 80 ℃ to constant weight. Industrial analysis of coal (moisture, ash, volatiles and fixed carbon) reference GB/T212-2008; the total calorific value of the coal is determined by a 5E-C5508 calorimeter (Hainan Kaiyuan Changsha in China) according to GB/TT 13-2008; vitrinite reflectance of coal was determined with reference to GB/T6948-2008 using a Leitz Orthoplan microscope (Solms, Germany) photometer. Through measurement, the coal contains 9.52 percent of water, 29.5 percent of ash, 44.04 percent of volatile matter, 26.46 percent of fixed carbon, 18.63MJ/kg of heat value of the coal and 0.39 percent of vitrinite reflectance. Referring to the classification standard of coal in coal seams in China (GB/T17607-1998), the coal sample used in the research is medium-ash high-order lignite.
Example 1
1) Nitric acid oxidation pretreatment of coal samples
Carrying out oxidation pretreatment on the lignite sample by using nitric acid with the concentration of 4N; the pretreated lignite samples were prepared by mixing lignite samples in a proportion of 1g per 25ml of the solution and carrying out an oxidation-reduction reaction. The redox reaction was carried out in a 50ml glass vial at 30 ℃ for 48 h. After the redox reaction, centrifuge at 7000rpm for 10min, wash the residual coal sample with deionized water until the wash solution is colorless, pH greater than 5. Then drying in an oven at 80 ℃ for 36 h.
2) Preparation of cell-free filtrates (CFFs)
The Penicillium MJ51MJ51(Penicillium camemberti, preservation number: CGMCC No.18120) is inoculated in PDA culture medium, cultured at 28 ℃ for 6 days, added with 0.1% physiological saline to form spore suspension, and observed by a blood cell counting plate under a microscope. 2.5mL (1X 10)7Spores) spore suspension was inoculated into a 250mL triangular flask containing 50mL of liquid medium having the following composition (g/L): sucrose 15.0, KNO33.0、KH2PO41.0、Na2HPO40.5，MgSO4·7H2O0.5, pH06.0. After inoculation, the cells were incubated at 160rpm and 28 ℃ for 60 h. Filtering the fungus suspension culture solution to separate into mycelium and extracellular fluid, collecting the extracellular fluid, measuring pH value, and filtering the supernatant with 0.22 μm membrane filter to obtain cell-free filtrate CFFs.
Example 2
The cell-free filtrate CFFs obtained in example 1 and the pretreated lignite sample were mixed, the coal loading was 4%, and the mixture was dissolved at 60 ℃ for 6 hours to obtain a solution, and the concentration of water-soluble humic acid was determined by a gravimetric method, and the determination result showed that the concentration of humic acid was 29.62 g/L.
Example 3
The cell-free filtrate CFFs obtained in example 1 and the pretreated lignite sample were mixed with a coal load of 4% and were dissolved at 60 ℃ for 10h, in contrast to example 1, the concentration of nitric acid in step 1) was 2N. The determination result shows that the concentration of the humic acid is 17.52 g/L.
Example 4
The cell-free filtrate CFFs obtained in example 1 and the pretreated lignite sample were mixed, the coal loading was 3%, and the mixture was dissolved at 75 ℃ for 8 hours. The determination result shows that the concentration of the humic acid is 30.68 g/L.
Example 5
The cell-free filtrate CFFs obtained in example 1 and the pretreated lignite sample were mixed with a coal load of 3% and were dissolved at 75 ℃ for 8h, unlike example 1, the concentration of nitric acid in step 1) was 6N. The determination result shows that the concentration of the humic acid is 32.29 g/L.
Example 6
The cell-free filtrate CFFs obtained in example 1 and the pretreated lignite sample were mixed, the coal loading was 5%, and the mixture was dissolved at 75 ℃ for 6 hours. The determination result shows that the concentration of humic acid is 29.28 g/L.
Example 7
The cell-free filtrate CFFs obtained in example 1 and the pretreated lignite sample were mixed, the coal loading was 4%, and the mixture was dissolved at 75 ℃ for 8 hours. The determination result shows that the concentration of the humic acid is 40.01 g/L.
Example 8
The cell-free filtrate CFFs obtained in example 1 and the pretreated lignite sample were mixed with a coal load of 4% and were dissolved at 75 ℃ for 10h, unlike example 1, the concentration of nitric acid in step 1) was 6N. The determination result shows that the concentration of humic acid is 32.32/L.
Example 9
The cell-free filtrate CFFs obtained in example 1 and the pretreated lignite sample were mixed with a coal load of 5% and were dissolved at 75 ℃ for 8h, unlike example 1, the concentration of nitric acid in step 1) was 6N. The determination result shows that the concentration of humic acid is 29.95/L.
Example 10
The cell-free filtrate CFFs obtained in example 1 and the pretreated lignite sample were mixed, with a coal loading of 4%, and dissolved at 90 ℃ for 6 hours. The determination result shows that the concentration of the humic acid is 33.18 g/L.
Example 11
The cell-free filtrate CFFs obtained in example 1 and the pretreated lignite sample were mixed, the coal loading was 3%, and the mixture was dissolved at 60 ℃ for 8 hours. The determination result shows that the concentration of the humic acid is 30.15 g/L.
Example 12
The cell-free filtrate CFFs obtained in example 1 and the pretreated lignite sample were mixed with a coal load of 4% and were dissolved at 75 ℃ for 6h, in contrast to example 1, the concentration of nitric acid in step 1) was 2N. The determination result shows that the concentration of the humic acid is 16.82/L.
Example 13
The cell-free filtrate CFFs obtained in example 1 and the pretreated lignite sample were mixed, the coal loading was 4%, and the mixture was dissolved at 90 ℃ for 10 hours. The determination result shows that the concentration of the humic acid is 38.05 g/L.
Example 14
The cell-free filtrate CFFs obtained in example 1 and the pretreated lignite sample were mixed, the coal loading was 4%, and the mixture was dissolved at 60 ℃ for 10 hours. The determination result shows that the concentration of the humic acid is 32.74 g/L.
Test example 3
Quality assessment of humic acid products
Coal dissolution was carried out by the method in example 7, and after the coal dissolution, the supernatant was filtered through Whatman No.1 filter paper. In order to conveniently evaluate various characterization indexes of the humic acid solution, the humic acid is separated by adopting an acid precipitation method, and the humic acid can be directly used without further acid precipitation in the application process of the humic acid. The specific operation steps are as follows: adjusting the pH of the filtrate to 1.8 by using 6.0M HCl, precipitating for 24h, centrifuging at 10000rpm for 5min, and collecting precipitated humic acid. Humic acid was washed three times with distilled water, dried at 60 ℃ and stored at 4 ℃ for further analysis. The control was humic acid extracted using 0.1M sodium hydroxide solution instead of cell free filtrate, as was the case with the other methods.
1. Elemental analysis of humic acid
The elemental composition of humic acid was measured with a Vario-EL elemental analyzer (germany), and the oxygen content was calculated from the difference. The E4/E6 values were determined using a SPECORD 200Analytik UV-visible spectrophotometer (Jena, Germany). The sample was dissolved in 0.05M NaHCO at pH 8.43The concentration of the solution was 20 mg/LHA. The resulting solution was centrifuged at 6,000rpm for 4min at room temperature to remove insoluble residues. Sodium bicarbonate buffer served as a blank.
See table 1 for results.
TABLE 1 comparison of chemical characteristics of different humic acids
Note: and represent significant differences in p <0.05, p <0.01 levels.
2. Humic acid flocculation limit (flocculation limit) is a measure of the ability of reacting humic acids to withstand electrolyte flocculation in the soluble state. Namely, electrolyte (0.1mol/L CaCl) is added when 0.02 percent humic acid alkali liquor is condensed within 1 hour2) Minimum milliequivalent number of (c). The method comprises the following specific steps: a series of dry test tubes were prepared and 2.5ml of CaCl of different concentrations were added2And (3) solution. 5ml of 0.02% HA solution was added to each tube. Covering the ground cap, shaking up, and standing for 1 h. The tubes were observed for the onset of precipitation (flocculation), and the flocculation limit was calculated as: flocculation limit (mmol/L) ═ M · V × 1000/5 ═ 200 MV; in the formula: M-CaCl2Concentration, here 0.1 mol/L; v-minimum CaCl at which the condensation limit is reached2Amount added (ml). The results are shown in Table 1. As can be seen from Table 1, the nitrogen and oxygen contents of humic acid extracted from lignite by the fungus cell-free filtrate are significantly increased, the sulfur and carbon contents are significantly reduced, and the H/C, O/C, flocculation limit value and E4/E6 value are significantly or extremely significantly increased (p) compared with the humic acid extracted from lignite by a chemical method<0.05，p<0.01). The higher the H/C and E4/E6 ratios, the lower the aromaticity and molecular mass. The increase of nitrogen content and the reduction of molecular weight of humic acid are beneficial to the absorption and utilization of plants, and the increase of the deflocculation capacity is beneficial to the application of the humic acid as liquid fertilizer.
3. Structural characterization of humic acid
Using FTIR and13C-qNMR analyzed the structure and functional group distribution of HA. To obtain FTIR spectra, a 2mg HA sample was mixed well with 200mg dry KBr and pressed into a tray using an IS10 FTIR spectrometer (Nicolet instruments Co., USA) at 4000400cm-1FTIR scanning of the sample is carried out in the range. Of Has sample13C nuclear magnetic resonance spectra were obtained using an AVANCE III 500MHz spectrometer (Bruker, Karlsruhe, German). For quantification, the spectra were divided into different chemical shift regions assigned to specific carbon groups. The results are shown in FIG. 1 and Table 2.
TABLE 2 results of carbon distribution in humic acid samples determined by solid-state CP/MAS 13C NMR
As can be seen from FIG. 1a, the two humic acids have similar FTIR spectra. The consensus peaks include: 3500-3300cm-1(O-H stretch in alcohol and phenol), 1720cm-1(carboxyl and carbonyl), 1380-1480cm-1(deformation vibrations of methylene and methyl groups) and 1035cm-1 (C-O stretch of polysaccharide component). The spectrum of the HA-CFFs sample was 1617cm compared to the HA control group-1And 911cm-1Contains no stretching and aromatic ring C-H bending absorption bands of C ═ C groups in the aromatic ring, which may be associated with degradation of the aromatic ring; while HA-CFFs have an absorption band at 1538cm-1, which is caused by C ═ N stretching and N — H deformation, the increase in nitrogen content in HA-CFF may be related to the increase in this functional group. FIG. 1b and Table 2 show that the methoxy, carboxyl and keto carbons in HA-CFFs are increased, the aliphatic carbon is increased and the aromatic carbon is decreased compared to the HA control. Since carboxyl and methoxy play an important role in promoting plant growth, this result suggests that HA-CFFs may have stronger physiological activity.
4. Biological activity of humic acid
To validate the effect of biological activity of HAs on corn growth, two treatments were established, three replicates of each treatment: (1) 0.03 kg of HA-control was applied-1(2) applying HA-CFFs 0.03gkg-1. The pots (20 cm. times.25 cm in size) were filled with 7.5kg of air-dried soil, and were arranged randomly. All treatments used the same amount of N, P2O5、K2O: 0.15, 0.1 and 0.12g kg respectively-1. Three corn seeds were sown per pot. After 30 days, the stem thickness, plant height, and aerial and underground biomass of each treated plant were measured. The data sets were tested for statistical significance using analysis of variance. p value<0.05 was considered statistically significant. The results are shown in Table 3.
TABLE 3 Effect of humic acids on corn growth
As can be seen from Table 3, the application of the HA-CFFs treatment significantly promoted the growth of the plants as compared to the application of HA-control, which was manifested by a significant increase in the stem thickness, plant height, underground biomass, overground biomass and whole plant biomass.
The foregoing is only a preferred embodiment of the present invention, and it should be noted that, for those skilled in the art, various modifications and decorations can be made without departing from the principle of the present invention, and these modifications and decorations should also be regarded as the protection scope of the present invention.
Claims (10)
1. Application of penicillium MJ51 extracellular fluid in preparation of water-soluble humic acid by dissolving lignite.
2. The use according to claim 2, wherein the method for preparing water-soluble humic acid by dissolving lignite comprises the following steps:
mixing the penicillium MJ51 extracellular fluid with oxidized lignite to obtain a mixed solution, and dissolving coal in the mixed solution at the temperature of 60-90 ℃ to obtain a dissolved solution, wherein the dissolved solution contains water-soluble humic acid.
3.The use according to claim 2, wherein the mass concentration of oxidized lignite in the mixed liquor is 3-5%.
4. The use according to claim 2, wherein the oxidized lignite is produced by a process comprising the steps of: mixing nitric acid and lignite, and then carrying out oxidation-reduction reaction;
and carrying out solid-liquid separation on the reaction feed liquid obtained by the oxidation-reduction reaction to obtain a solid component, wherein the solid component is oxidized lignite.
5. The use according to claim 4, wherein the temperature of the redox reaction is 25 to 30 ℃.
6. The use according to claim 4, wherein the nitric acid has an equivalent concentration of 4 to 6N.
7. The use according to claim 1 or 2, wherein the penicillium MJ51 extracellular fluid is prepared by a method comprising the steps of:
inoculating the spore suspension of the penicillium MJ51 into a liquid culture medium, and culturing to obtain a culture solution;
isolating the extracellular fluid of penicillium MJ51 from the culture medium;
the spore concentration of the penicillium MJ51 in the spore suspension is (1-9) multiplied by 107A plurality of; the volume ratio of the spore suspension to the culture medium is 5: (80-120).
8. The use according to claim 7, wherein the temperature of the culture is 25-30 ℃; the culture time is 55-65 h.
9. The use according to claim 7, wherein the liquid medium is water as solvent and comprises the following components in the following concentrations: 12-18 g/L, KNO of sucrose32～4g/L、KH2PO40.8～1.2g/L、Na2HPO40.3～0.8g/L、MgSO40.4-0.6 g/L; the pH value of the culture medium is 5.8-6.2.
10. Use according to claim 7, wherein the penicillium MJ51 has the deposit number: CGMCC No. 18120.
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| CN114574879A (en) * | 2022-03-02 | 2022-06-03 | 大连理工大学 | Method for improving yield of humic acid prepared by coal electrolysis by coating film on surface of coal particles |
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| CN101967072A (en) * | 2010-08-11 | 2011-02-09 | 西北农林科技大学 | Organic-inorganic compound fertilizer |
| CN110699266A (en) * | 2019-11-27 | 2020-01-17 | 河南农业大学 | Penicillium MJ51 and application thereof |
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| CN101967072A (en) * | 2010-08-11 | 2011-02-09 | 西北农林科技大学 | Organic-inorganic compound fertilizer |
| CN110699266A (en) * | 2019-11-27 | 2020-01-17 | 河南农业大学 | Penicillium MJ51 and application thereof |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN114574879A (en) * | 2022-03-02 | 2022-06-03 | 大连理工大学 | Method for improving yield of humic acid prepared by coal electrolysis by coating film on surface of coal particles |
| CN114574879B (en) * | 2022-03-02 | 2023-09-22 | 大连理工大学 | Method for improving yield of humic acid prepared by coal electrolysis through coal particle surface coating |
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